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A SIMPLE AND RAPID MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY METHOD TO SCREEN FISH PLASMA SAMPLES FOR ESTROGEN-RESPONSIVE BIOMARKERS
SALINAS, K., M. J. HEMMER, P. S. HARRIS, AND C. C. WALKER. A SIMPLE AND RAPID MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY METHOD TO SCREEN FISH PLASMA SAMPLES FOR ESTROGEN-RESPONSIVE BIOMARKERS. ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY. Society of Environmental Toxicology and Chemistry, Pensacola, FL, 27(5):1175-1183, (2008).
Description and evaluation of a simple rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers.
In this study, we describe and evaluate the performance of a simple and rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers using matrix assisted laster desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) couopled with a short term fish assay. Adult male sheepshead minnows (Cyprinodon variegatus) were placed into aquaria consisting of duplicate tanks for vehicle control and the following estrogen agonist treatments: 0.00713, 0.0163, 0.04, 0.05, 0.1305, 0.222, 0.399 and 0.906 ug 17B-esstradiol/l; 100 ug 4-tert-pentylephenol /l; 6 and 12 ug methoxychlor/l; and 120.5 and 1079.5 ug bisphenol-A/l. Treatments with 80 ug chlorpyrifos/l and 0.6 ug endosulfan/l served as non-estrogenic negative controls. Test concentrations were maintained using an intermittent flow-through dosing apparatus, and fish were sampled at seven or 10 days. Plasma was obtained from individuals, diluted and applied to an inert gold array and analyzed. Multiple protein peaks, ranging from 2.9 to 12.9 KSa, were identified as markers of estrogenic effects when comparing estrogen-treated and control fish using interpercentile reference values. A binary classificiton tree model was constructed from plasma protein profiles of the vehicle control and 0.222 ug/l 17B-estradiol treatments and used to evaluate all samples. Treatments with estrogen agonists 17B-estradiol, 4-tert-pentylphenol, methoxychlor and bisphenol-A generated reproducible diagnostic biomarkers based on the presence of specific estrogen-responsive plasma proteins. The controls and non-estrogenic compounds chlorpyrifos and endosulfan did not produce this estrogen responsive protein profile. An average measured NOEC for 17B-estradiol at 0.04 ug/l was estimated from concentration response exposures. MALDI-TOF-MS is a sensitive adn specific tool for protein expression profiling and can be used to screen chemicals for mode of action in ecotoxicological studies.
Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL HEALTH AND ENVIRONMENTAL EFFECTS RESEARCH LABORATORY
GULF ECOLOGY DIVISION
BIOLOGICAL EFFECTS AND POPULATION RESPONSE BRANCH